GSM7977975: RNA lungs, Control, rep2; Mus musculus; RNA-Seq
Lungs from E17.5 mice were harvested, minced, and incubated in RPMI media containing Liberase TM (Roche, 50ug/ml) and DNAse I (MilliporeSigma, 100ug/ml) at 37C for 30min. Tissues were dissociated with GentleMACS cell dissociator, then subjected to RBC lysis buffer for 2min on ice, followed by an incubation with FcR blocking reagent (Miltenyi: 130-092-575) on ice for 10min, followed by an incubation with CD326 anti-Mouse-EpCAM microbeads (Miltenyi: 130-105-958) at 4C for 15min. Cells were subjected to positive selection with EpCAM+ microbeads in an LS column and collected in MACS buffer (Miltenyi). Cells were quantified manually via hemacytometer with trypan blue. RNA was isolated from selected cells using Qiagen's Rneasy Micro Plus kit following manufacturer's directions. Collected RNA was sent to Genewiz for polyA enrichment, library construction, and sequencing.